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Protease Activated Receptors and Arthritis

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Protease Activated Receptors and Arthritis International Journal of Molecular Sciences Review Protease Activated Receptors and Arthritis Flora Lucena and Jason J. McDougall * Departments of Pharmacology and Anesthesia, Pain Management & Perioperative Medicine, Dalhousie University, 5850 College Street, Halifax, NS B3H 4R2, Canada; fl[email protected] * Correspondence: [email protected] Abstract: The catabolic and destructive activity of serine proteases in arthritic joints is well known; however, these enzymes can also signal pain and inflammation in joints. For example, thrombin, trypsin, tryptase, and neutrophil elastase cleave the extracellular N-terminus of a family of G protein- coupled receptors and the remaining tethered ligand sequence then binds to the same receptor to initiate a series of molecular signalling processes. These protease activated receptors (PARs) pervade multiple tissues and cells throughout joints where they have the potential to regulate joint homeostasis. Overall, joint PARs contribute to pain, inflammation, and structural integrity by altering vascular reactivity, nociceptor sensitivity, and tissue remodelling. This review highlights the therapeutic potential of targeting PARs to alleviate the pain and destructive nature of elevated proteases in various arthritic conditions. Keywords: arthritis; inflammation; joint damage; proteases; pain 1. Introduction Musculoskeletal diseases comprise the most prevalent chronic pain conditions with Citation: Lucena, F.; McDougall, J.J. arthritides accounting for the majority of these disorders [1]. Although there are over Protease Activated Receptors and 100 different types of arthritis, the most commonly studied are inflammatory rheumatoid Arthritis. Int. J. Mol. Sci. 2021, 22, arthritis (RA) and degenerative osteoarthritis (OA). In RA, an individual’s immune system 9352. https://doi.org/10.3390/ is dysregulated and inflammatory cells begin to destroy the host joint tissues by releasing ijms22179352 chemical mediators into the joint. Autoantibodies such as rheumatoid factor and antibodies directed towards activated citrullinated proteins are also prominent features of RA which Academic Editor: Giovanni Tarantino contribute to the degenerative process and pain [2]. The RA synovium becomes hyperplas- tic and this pannus invades cartilage, bone, and menisci leading to altered biochemical Received: 21 July 2021 regulation and tissue damage. OA is characterized as an inappropriate healing response to Accepted: 26 August 2021 joint tissue injury whereby cartilage exhibits focal lesions, subchondral bone is dense and Published: 28 August 2021 fissured, osteophytes form, and the menisci can become calcified [3]. Intermittent synovitis occurs in some patients and joint inflammation is heterogeneous [4]. Pain and joint stiff- Publisher’s Note: MDPI stays neutral ness are common to all types of arthritis and limit an individual’s ability to move which with regard to jurisdictional claims in could put them at risk of other co-morbidities such as diabetes, obesity, or cardiovascular published maps and institutional affil- disease [5–7]. iations. The available treatments to manage joint diseases are currently very limited and have variable efficacy between patients. Pharmacological treatments for RA include the use of non-steroidal anti-inflammatory drugs (NSAIDs), corticosteroids, methotrexate, and biologics that inhibit cytokine reactivity [8]. With respect to OA, there are no disease Copyright: © 2021 by the authors. modifying drugs and treatments are restricted to symptom relief. Topical NSAIDs, ac- Licensee MDPI, Basel, Switzerland. etaminophen, serotonin-noradrenaline reuptake inhibitors, and opioids are typically used This article is an open access article to treat the disease [9]. All of the drug therapies have varying degrees of effectiveness and distributed under the terms and can produce undesirable side effects. The search for new drug targets that can control pain, conditions of the Creative Commons inflammation, and function is therefore an important area for ongoing arthritis research. Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ The chemical mediators responsible for generating joint disease are still being defined 4.0/). but are known to include prostaglandins, neuropeptides, and cytokines [10]. Emerging Int. J. Mol. Sci. 2021, 22, 9352. https://doi.org/10.3390/ijms22179352 https://www.mdpi.com/journal/ijms Int. J. Mol. Sci. 2021, 22, 9352 2 of 16 evidence also indicates that proteolytic enzymes can also signal pain and inflammation in joints by cleaving a specific receptor family known as the protease activated receptors (PARs; Table1). This group consists of four G-protein coupled receptors (PAR-1 to -4) that show a unique activation characteristic [11]. In the presence of a protease, a portion of the PAR extracellular N-terminus is cleaved which then exposes a new N-terminus sequence that acts as a tethered ligand which then binds to the second extracellular loop of the receptor [12]. This conformational change to the receptor leads to intracellular signalling that will vary depending on the receptor subtype, the cleaving protease, and the downstream pathway that becomes activated. In addition to receptor activation, some proteases can disarm or inactivate the PAR receptor by cleaving the N-terminus at an adjacent site [13]. More recently, a biased signalling of PARs has been extensively described, indicating the complexity of these receptors [14–16]. PARs have been identified on the nerve terminals of nociceptors in multiple tissues suggesting that this receptor family is involved in pain control. Co-localization and phar- macological interaction between PAR-2 and pro-algesic transient receptor potential (TRP) channels in the pancreas [17], bladder [18], and oral mucosa [19] indicate that there is functional coupling between PARs and other known pain-modulating receptors. Proteases released from immunocytes, endothelial cells, or are part of the coagulation cascade are known to cleave neuronal PARs and modulate pain neurotransmission [12]. Other studies on the skin, gut, and airways have shown that PAR activation can lead to the secondary release or pro-algesic neuropeptides such as substance P and calcitonin gene-related pep- tides [20–22]. These multiple lines of evidence assert that PARs are an attractive target for the management of miscellaneous chronic pain conditions. Table 1. Known cleaving proteases and synthetic activating peptides for each of the PARs and their role in arthritis. Synthetic Activating PAR Activating Proteases Effect in Joints Peptides Thrombin SFLLRN-NH2 Chondroprotection [23] Granzyme A TFLLRN-NH2 Bone repair [24] Plasmin Pannus formation [25] Activating protein C Anti-allodynic via an opioid mechanism [26] Trypsin Factor Xa PAR-1 Kallikrein- 4, 5, 6, 14 MMP-1 Cathepsin G Proatherocytin Pen C 13 Chymase Trypsin SLIGRL-NH2 Cartilage degeneration [27] Synovial hyperaemia and increased Mast cell tryptase FLIGRL-NH2 leukocyte trafficking [28] TRPV -dependent afferent sensitization and Factor Xa: Factor VIIa 1 pain [29,30] PAR-2 Acrosin Matriptase Serine 11D Trypsin Granzyme A Kallikrein-2, 4, 5, 6, 14 PAR-3 Thrombin N/A N/A Int. J. Mol. Sci. 2021, 22, 9352 3 of 16 Table 1. Cont. Synthetic Activating PAR Activating Proteases Effect in Joints Peptides Thrombin AYPGKF-NH2 Joint damage [31] Trypsin GYPGKF-NH2 Joint hyperaemia and oedema [32] Afferent sensitization and pain via mast cell Cathepsin G degranulation and bradykinin activation [32] PAR-4 Trypsin IV Mannin-binding SP-1 Plasmin Factor Xa Kallikrein-1, 14 C4a This review will outline the function of PARs and give an overview of the latest studies, implicating their role in joint diseases including RA and OA. 2. PAR-1 2.1. Receptor Pharmacology PAR-1 receptors are mainly activated by thrombin, but other mediators of the coagula- tion cascade can also cleave it. PAR-1 is ubiquitously found in endothelial cells, platelets, lungs, GI tract, immune cells, neurones and brain [11]. Following cleavage of PAR-1, there is activation of the G protein subunits G12/13,Gi, and Gq (Figure1A) [ 33–36]. Signalling via G12/13 leads stimulation of Rho guanine nucleotide exchange factor (RhoGEF), which in turn activates GTPase RhoA and is involved in cytoskeleton reorganization. Activation of Gi causes inhibition of adenylyl cyclase activity and hence a reduction in cyclic adeno- sine monophosphate (cAMP) production. Finally, Gq activation initiates the intracellular cascade starting with phospholipase-Cβ (CPLCβ) hydrolyzing phosphatidylinositol 4,5- bisphosphate (PIP2) to produce inositol triphosphate (IP3) and diacylglycerol (DAG). This leads to Ca2+ mobilization and an increase in the activity of protein kinase C (PKC) and other intracellular signalling enzymes. Different subunits of PKC facilitate the variety of downstream responses after PAR-1 activation [37–40]. A peculiar characteristic of PARs is that because its agonist is its own N-terminal sequence, there is no dissociation of the agonist from the receptor, so desensitization and termination mechanisms have to be tightly regulated. PAR-1 receptors are phosphorylated by G-protein coupled receptor kinases (GRKs) 3 or 5 and desensitized by β-arrestin binding, specifically β-arrestin 1, allowing the uncoupling of the G-proteins [12]. In contrast to other GPCRs, the β-arrestin associated with PAR-1 participates only in desensitization of the receptor, but not internalization [41]. Receptor internalization is primarily regulated by clathrin and dynamin activity.
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